结肠癌细胞系SW620中ALDH1^(br)细胞特性的研究

Preliminary study on the cellular properties of ALDH1^(br) cells in colonic cancer cell line SW620

目的检测结肠癌细胞系SW620中是否存在肿瘤干细胞样细胞。方法按照ATCC标准培养SW620细胞,用AL-DEFLUOR试剂盒联合荧光激活细胞分类术(FACS)分选ALDH1br细胞,无血清培养2周后再次进行FACS检测,观察ALDH1br细胞自我更新及增殖情况;检测ALDH1br细胞血清诱导1周后分化标志物CK20含量、ALDH1活性及耐药蛋白ABCG2含量的变化;Transwell小室实验检测ALDH1br细胞的迁移能力。结果经FACS检测,SW620中ALDH1br细胞比例为3.59%;无血清培养2周后,ALDH1br细胞增加至43.0%±2.2%,且不对称性分裂为ALDH1br细胞和ALDH1-细胞;经血清诱导1周后,ALDH1br细胞分化标志物CK20含量明显增高(0.64±0.040、.01±0.02,P<0.05),且ALDH1br细胞比例下降(1.73%);分选后ALDH1br细胞的耐药蛋白ABCG2含量也明显增加(0.27±0.03、0.90±0.05,P<0.05);ALDH1br细胞比普通的SW620细胞具有更强的迁移能力(细胞穿孔数分别为58.5±3.6、9.4±2.1,P<0.05)。结论运用ALDEFLUOR试剂联合FACS从结肠癌细胞系SW620中分选出的ALDH1br细胞具有肿瘤干细胞样特征。

Objective To isolate the ALDH1br cells from SW620 cell line,for the study of its stem cell like property in order to prove whether tumor stem cell like cells existed in SW620 cell line.Methods The SW620 cell line was cultured according to the ATCC standard,and the ALDH1br cells were separated with ALDEFLOUR reagent kit combined with FACS,and then cultivated serum-free for 2 weeks.The ALDH1br cells were detected once again with FACS to observe their self-renewal and proliferation.After serum-inducing for one week,the change in cell differentiation marker CK20 was examined by RT-PCR and Western-blotting,that of ALDH1 with RT-PCR and FACS,and that of the drug-fast protein ABCG2 with RT-PCR and Western-blotting.The migration ability of ALDH1br cells was detected by Transwell experiment.Results The content of ALDH1br cells in SW620 cell line was 3.59%.After being cultivated for two weeks in SFM,the proliferation and asymmetrical division abilities of ALDH1br cells in vitro were enhanced greatly;after serum-inducing,the proportion of cell differentiation marker CK20 increased obviously,while the contents of ALDH1 decreased markedly.After separation,the contents of drug-fast protein ABCG2 in ALDH1br cells increased obviously,and then decreased by serum-inducing.The ALDH1br cells showed greater migration ability than the SW620 cells.Conclusion ALDH1br cells,separated from colonic cancer cell line SW620 with ALDEFLUOR reagent combined with FACS,have the characteristics of stem cell.